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Affinity chromatography purification of diphtheria toxin
Author(s) -
Cukor George,
Readio Josephine D.,
Kuchler Robert J.
Publication year - 1974
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260160706
Subject(s) - chromatography , diphtheria toxin , elution , chemistry , toxin , antitoxin , sepharose , phosphate buffered saline , affinity chromatography , biochemistry , enzyme
NAD was covalently linked to Sepharose‐4B using a 6 carbon spacer. Sterile, dialyzed spent culture medium containing 100 Lf/ml of diphtheria toxin or material concentrated by (NH 4 ) 2 SO 4 precipitation containing 1500 Lf/ml, was chromatographed on a column of NAD–Sepharose. Ultraviolet absorbing material which did not flocculate with diphtheria antitoxin was eluted with 0.02 M phosphate buffer. When the elation buffer was changed to one containing 0.5 M NaCl, purified toxin was eluted off the column.